Figure 5.

miR-363 directly targeted Snail1 in RCC cells. A: To clone 3’-UTR sequences of wild type and mutant Snail1 pGL3-promoter luciferase vector was used. To justify the putative binding-sites between 3’-UTR and miR-363, matched base pairs were presented with solid lines and unmatched with cross lines. B: miRNA control (miR-NC), or miR363 mimic, Snail1-Mut reporter and wild-type (WT) Snail1 can be used for co-transfection with OSRC-2 and SW839 cells. Then, 48 hours post-transfection the luciferase activity was detected; C: The validation of the interaction between Snail1 and miR-363 in OSRC-2 cells and SW839 cells were conducted with RNA pull-down assays; D: Ago2 or IgG antibody were used in RNA-immunoprecipitation assays to investigate the enrichment of snail1 and miR-363 in Ago2 or IgG immunoprecipitation complexes of OSRC-2 cells and SW839 cells. The input group is a positive control; E: To detect the expression level of Snail1 in RCC cells after overexpressing miR-363; western blot was performed. F: Western blot was used for the measurement of Snail expression in RCC cells after knocked out of miR-363; G: Expression level of a snail after treatment with propofol was measured in SW839 and OSRC-2 cells. Data are presented in mean ± S.E.M. ***P<0.001, while comparing with control (ANOVA and post hoc test).